Home LiteratureArticle Details
PMID: 1903539 Published · ppublish English Journal Article

Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 10 ·1991-05-15 ·Pages 4328-32

Toledano MB, Leonard WJ

Abstract

NF-kappa B is a widely used regulator of inducible and tissue-specific gene control. In the cytosol, when complexed to an inhibitory molecule, I kappa B, NF-kappa B is in an inactive form and cannot bind DNA. Activation of cells with appropriate stimuli results in the dissociation of NF-kappa B from I kappa B and its translocation to the nucleus as an active binding protein. We now demonstrate that NF-kappa B binding in vitro can be inhibited by agents that modify free sulfhydryls. Binding is eliminated after treatment with N-ethylmaleimide, an alkylating agent, and diamide, an oxidizing agent. The diamide effect can be reversed by 2-mercaptoethanol. Further, 2-mercaptoethanol acts synergistically with deoxycholate plus Nonidet P-40 in converting inactive cytosolic NF-kappa B to an active DNA-binding form. It is therefore possible that modulation of the redox state of NF-kappa B could represent a post-translational control mechanism for this factor.

MeSH Terms
Cell Nucleus/metabolism Cytosol/metabolism DNA/metabolism DNA-Binding Proteins/metabolism Deoxycholic Acid/pharmacology Diamide/pharmacology Drug Synergism Ethylmaleimide/pharmacology Humans Mercaptoethanol/pharmacology NF-kappa B/chemistry,metabolism Nuclear Proteins/metabolism Octoxynol Oxidation-Reduction Polyethylene Glycols/pharmacology Proto-Oncogene Proteins c-jun Serum Response Factor Sulfhydryl Compounds/chemistry Transcription Factors/metabolism Tumor Cells, Cultured
Chemicals
DNA-Binding Proteins NF-kappa B Nuclear Proteins Proto-Oncogene Proteins c-jun Serum Response Factor Sulfhydryl Compounds Transcription Factors Deoxycholic Acid Diamide Polyethylene Glycols Mercaptoethanol Octoxynol DNA Nonidet P-40 Ethylmaleimide
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Toledano M B
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesa, MD 20892.
Leonard W J
References (40)
40 references, click to expand
  1. NF-kappa B protein purification from bovine spleen: nucleotide stimulation and binding site specificity.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):8825-9 PMID: 2848241
  2. Glutathione. 13. Mechanism of thiol oxidation by diazenedicarboxylic acid derivatives.
    J Am Chem Soc. 1976 May 12;98(10):3001-7 PMID: 1262629
  3. Oligonucleotide that binds nuclear factor NF-kappa B acts as a lymphoid-specific and inducible enhancer element.
    Proc Natl Acad Sci U S A. 1988 Mar;85(5):1482-6 PMID: 3125549
  4. Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-kappa B transcription factor.
    Cell. 1988 Apr 22;53(2):211-7 PMID: 3129195
  5. The 'CC.Ar.GG' box. A protein-binding site common to transcription-regulatory regions of the cardiac actin, c-fos and interleukin-2 receptor genes.
    Eur J Biochem. 1988 May 2;173(3):507-15 PMID: 3131140
  6. I kappa B: a specific inhibitor of the NF-kappa B transcription factor.
    Science. 1988 Oct 28;242(4878):540-6 PMID: 3140380
  7. Identification and purification of a polypeptide that binds to the c-fos serum response element.
    EMBO J. 1987 Sep;6(9):2711-7 PMID: 3119326
  8. c-Jun dimerizes with itself and with c-Fos, forming complexes of different DNA binding affinities.
    Cell. 1988 Dec 2;55(5):917-24 PMID: 3142692
  9. DNA binding activities of three murine Jun proteins: stimulation by Fos.
    Cell. 1988 Dec 2;55(5):907-15 PMID: 3142691
  10. Functionally distinct NF-kappa B binding sites in the immunoglobulin kappa and IL-2 receptor alpha chain genes.
    Science. 1989 Apr 28;244(4903):466-9 PMID: 2497520
  11. A 65-kappaD subunit of active NF-kappaB is required for inhibition of NF-kappaB by I kappaB.
    Genes Dev. 1989 Nov;3(11):1689-98 PMID: 2691328
  12. cis-trans models for post-transcriptional gene regulation.
    Science. 1989 Nov 17;246(4932):870-2 PMID: 2683086
  13. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control.
    Cell. 1989 Jul 28;58(2):227-9 PMID: 2665943
  14. ATL-derived factor (ADF), an IL-2 receptor/Tac inducer homologous to thioredoxin; possible involvement of dithiol-reduction in the IL-2 receptor induction.
    EMBO J. 1989 Mar;8(3):757-64 PMID: 2785919
  15. Irreversible inhibition of v-src tyrosine kinase activity by herbimycin A and its abrogation by sulfhydryl compounds.
    Biochem Biophys Res Commun. 1989 Sep 15;163(2):803-9 PMID: 2783121
  16. Oxidation-reduction and the molecular mechanism of a regulatory RNA-protein interaction.
    Science. 1989 Apr 21;244(4902):357-9 PMID: 2711187
  17. Identification of a putative regulator of early T cell activation genes.
    Science. 1988 Jul 8;241(4862):202-5 PMID: 3260404
  18. Isolation and properties of cDNA clones encoding SRF, a transcription factor that binds to the c-fos serum response element.
    Cell. 1988 Dec 23;55(6):989-1003 PMID: 3203386
  19. Activation of the interleukin-2 receptor alpha gene: regulatory role for DNA-protein interactions flanking the kappa B enhancer.
    New Biol. 1989 Oct;1(1):83-92 PMID: 2518691
  20. Adult T-cell leukemia-derived factor/thioredoxin, produced by both human T-lymphotropic virus type I- and Epstein-Barr virus-transformed lymphocytes, acts as an autocrine growth factor and synergizes with interleukin 1 and interleukin 2.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8282-6 PMID: 2172979
  21. Intracellular thiols regulate activation of nuclear factor kappa B and transcription of human immunodeficiency virus.
    Proc Natl Acad Sci U S A. 1990 Dec;87(24):9943-7 PMID: 2263644
  22. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor.
    Cell. 1987 Jun 19;49(6):729-39 PMID: 3034432
  23. Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB.
    Nature. 1988 Aug 11;334(6182):494-8 PMID: 2900470
  24. Identification and purification of a human immunoglobulin-enhancer-binding protein (NF-kappa B) that activates transcription from a human immunodeficiency virus type 1 promoter in vitro.
    Proc Natl Acad Sci U S A. 1988 Jul;85(13):4700-4 PMID: 3133660
  25. The same inducible nuclear proteins regulates mitogen activation of both the interleukin-2 receptor-alpha gene and type 1 HIV.
    Cell. 1988 Jun 3;53(5):827-36 PMID: 2836068
  26. Human proto-oncogene c-jun encodes a DNA binding protein with structural and functional properties of transcription factor AP-1.
    Science. 1987 Dec 4;238(4832):1386-92 PMID: 2825349
  27. Formation of disulfides with diamide.
    Methods Enzymol. 1987;143:264-70 PMID: 3657543
  28. Xenopus cytoskeletal actin and human c-fos gene promoters share a conserved protein-binding site.
    EMBO J. 1987 Mar;6(3):667-73 PMID: 3582369
  29. Mutation of the c-fos gene dyad symmetry element inhibits serum inducibility of transcription in vivo and the nuclear regulatory factor binding in vitro.
    Mol Cell Biol. 1987 Mar;7(3):1217-25 PMID: 3561415
  30. Cloning of the p50 DNA binding subunit of NF-kappa B: homology to rel and dorsal.
    Cell. 1990 Sep 7;62(5):1019-29 PMID: 2203532
  31. The DNA binding subunit of NF-kappa B is identical to factor KBF1 and homologous to the rel oncogene product.
    Cell. 1990 Sep 7;62(5):1007-18 PMID: 2203531
  32. Redox regulation of fos and jun DNA-binding activity in vitro.
    Science. 1990 Sep 7;249(4973):1157-61 PMID: 2118682
  33. Glutathione regulates activation-dependent DNA synthesis in highly purified normal human T lymphocytes stimulated via the CD2 and CD3 antigens.
    Proc Natl Acad Sci U S A. 1990 May;87(9):3343-7 PMID: 1970635
  34. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation.
    Science. 1990 Apr 13;248(4952):189-94 PMID: 2183352
  35. The same target sequences are differentially important for activation of the interleukin 2 receptor alpha-chain gene in two distinct T-cell lines.
    Proc Natl Acad Sci U S A. 1990 Mar;87(5):1830-4 PMID: 2308942
  36. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  37. Complete nucleotide sequence of a human c-onc gene: deduced amino acid sequence of the human c-fos protein.
    Proc Natl Acad Sci U S A. 1983 Jun;80(11):3183-7 PMID: 6574479
  38. Nonspecific activation of murine lymphocytes. V. Role of cellular collaboration between T and B lymphocytes in the proliferative and polyclonal response to 2-mercaptoethanol.
    J Immunol. 1979 Apr;122(4):1433-9 PMID: 312842
  39. Effects of glutathione-oxidizing agents on microtubule assembly and microtubule-dependent surface properties of human neutrophils.
    J Cell Biol. 1976 Dec;71(3):921-32 PMID: 993272
  40. An inducible transcription factor activates expression of human immunodeficiency virus in T cells.
    Nature. 1987 Apr 16-22;326(6114):711-3 PMID: 3031512
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.10.4328
Published
1991-05-15
Pages
4328-32
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51652
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.88.10.4328 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2